Abstract
Fertilized embryo sacs of Zea mays L. surrounded by a few layers of nucellar cells were cultured in vitro. Primary expiants contained zygotes or twocelled proembryos. Embryos of various sizes and shapes were isolated from 12–48% of explants after two weeks of culture in hormone-free media supplemented with 6–12% of sucrose. Many embryos were at the transition or proembryo stages whilst the rest were either differentiated, with a scutellum, a coleoptile and a shoot apex, or had a deformed apical part. Organogenesis started in 36–89% of embryos cultured on a semisolid medium supplemented with coconut water. Most of the embryos formed only roots but up to 9% of embryos regenerated into plants. This simple method leads the way to plant regeneration from in-vitro-manipulated zygotes or proembryos of maize.
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Abbreviations
- NBM:
-
medium composed of N6 macronutrients, B5 micronutrients and MS vitamins
References
Abbe, E.C., Stein, O.L. (1954) The growth of the shoot apex in maize: embryogeny. Am. J. Bot. 41, 285–293
Chu, C.C. (1978) The N6 medium and its application to anther culture of cereal crops. In: Proc. Symp. Plant Tissue Culture, pp. 43–50, Chinese Acad. Sci., ed. Science Press, Peking
Clark, J.K., Sheridan, W.F. (1991) Isolation and characterization of 51 embryo-specific mutations of maize. Plant Cell 3, 935–951
Comeau, A., Nadeau, P., Plourde, A., Simard, R., Maës, O., Kelly, S., Harper, L., Lettre, J., Landry, B., St-Pierre, C.A. (1992) Media for the in ovulo culture of proembryos of wheat and wheat-derived interspecific hybrids or haploids. Plant Sci. 81, 117–125
Custers, J.B.M., Bergervoet, J.H.W. (1990) In vitro culture of embryos of Cucumis spp.: heart-stage embryos have a higher ability of direct plant formation than advanced-stage embryos. Sex. Plant Reprod. 3, 152–159
Gamborg, O., Miller, R., Ojima, K. (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp. Cell Res. 150, 151–158
Johri, B.M., Rao, P.S. (1984) Experimental embryology. In: Embryology of angiosperms, pp. 735–803, Johri, B.M., ed. Springer, Berlin Heidelberg New York Tokyo
Kapoor, M. (1959) Influence of growth substances on the ovules of Zephyranthes. Phytomorphology 9, 313–315
Kranz, E., Bautor, J., Lörz, H. (1991) In vitro fertilization of single, isolated gametes of maize mediated by electrofusion. Sex. Plant Reprod. 4, 12–16
Matthys-Rochon, E. (1992) In vitro fertilization in flowering plants. In: Reproductive biology and plant breeding, pp. 197–204, Dattée, Y., Dumas, C., Gallais, A., eds. Springer, Berlin Heidelberg New York
Murashige, T., Skoog F. (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15, 473–497
Nakajima, T., Wakizuka, T. (1977) Ovule culture of Petunia hybrida Vilm. In: Interspecific hybridization in plant breeding, Proc. 8th Congr. Eucarpia, pp. 253–258, Sanchez-Monge, E., Garcia-Olmedo, F., eds. Universidad Polytecnica, Madrid
Neuhaus, G., Lusardi, M.C. (1991) Maize somatic and zygotic embryos as target for maize transformation. In: Abstr. 1st Maize European Network Meeting, p. 32, Gargnano, Italy
Raghavan, V. (1986) Embryogenesis in angiosperms. A developmental and experimental study. Cambridge University Press, Cambridge London New York New Rochelle Melbourne Sydney
Raghavan, V., Srivastava, P.S. (1982) Embryo culture. In: Experimental embryology of vascular plants, pp. 195–230, Johri, B.M., ed. Springer, New York
Rangan, T.S. (1982) Ovary, ovule, and nucellus culture. In: Experimental embryology of vascular plants, pp. 105–129, Johri, B.M., ed. Springer, New York
Schel, J.H.N., Kieft, H. (1986) An ultrastructural study of embryo and endosperm development during in vitro culture of maize ovaries (Zea mays). Can. J. Bot. 64, 2227–2238
Töpfer, R., Steinbiss, H.H. (1985) Plant regeneration from cultured fertilized barley ovules. Plant Sci. 41, 49–54
Van Lammeren, A.A.M. (1986) Developmental morphology and cytology of the young maize embryos (Zea mays L.). Acta Bot. Neerl. 35, 169–188
Van Lammeren, A.A.M. (1988) Observations on the structural development of immature maize embryos (Zea mays L.) during in vitro culture in the presence or absence of 2,4-D. Acta Bot. Neerl. 37, 49–61
Young, B.A., Sherwood, R.T., Bashaw, E.C. (1979) Cleared-pistil and thick-sectioning techniques for detecting aposporous apomixis in grasses. Can. J. Bot. 57, 1668–1672
Zenkteler, M., Nitzsche, W. (1990) In vitro culture of wheat ovules. In: Biotechnology in agriculture and forestry, vol. 13: Wheat, pp. 262–268, Bajaj, Y.P.S., ed. Springer, Berlin Heidelberg
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This research was supported by an I.N.R.A. post-doctoral fellowship. The authors thank R. Blanc for donor plant culture, Dr. M. Cock (Reconnaissance Cellulaire et Amélioration des Plantes, Université Lyon 1) for correction of the English and P. Audenis for micrograph development.
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Mòl, R., Matthys-Rochon, E. & Dumas, C. In-vitro culture of fertilized embryo sacs of maize: Zygotes and two-celled proembryos can develop into plants. Planta 189, 213–217 (1993). https://doi.org/10.1007/BF00195079
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DOI: https://doi.org/10.1007/BF00195079